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Recognizes 4-HNE-lysine, 4-HNE-histidine and 4-HNE cysteine adduct. Has much higher affinity for the 4-HNE-histidine adduct than 4-HNE-lysine or 4-HNE cysteine adduct. Shows almost negligible reactivity with proteins that were treated with other aldehydes, such as 2-nonenal, 2-hexenal, 1-hexanal, 4-hydroxy-2-hexenal, formaldehyde, or glutaraldehyde.

Reactive oxygen species (ROS) are involved in lipid peroxidation inside the living bodies, and membrane lipids are one of the major targets of ROS. During the peroxidation process, a variery of aldehydes are formed. 4-hydroxy-2-nonenal (4-HNE) is an alpha, beta unsaturated aldehyde that can be formed by peroxidation of omega-6 unsaturated fatty acids such as linoleic acid and arachidonic acid. Especially, 4-HNE formed inside living body is reported to originate from phospholipid-bound arachidonic acid. 4-HNE may be one of the most reliable biomarker of lipid peroxidation.